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通过输送稳定的CRISPR-Cas9核糖蛋白的脂质纳米粒子来编辑肺和肝脏

Kai Chen^1,2, Hesong Han^2,3, Sheng Zhao^2,3

¹Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, USA.

Nature biotechnology

|October 16, 2024

概括

在脂质纳米粒子 (LNP) 中设计的Cas9变体实现了100倍以上的基因组编辑效率. 聚类定期间隔短时间的巴林德罗姆重复 (CRISPR) 交付的这一突破显示出对体内基因治疗的前景.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
基因编辑技术的技术

背景情况:

脂质纳米颗粒 (LNP) 输送集群定期间隔的短时间palindromic重复 (CRISPR) 核糖蛋白 (RNP) 是一个有前途的策略 in vivo基因组编辑.
通过可靠的RNP-LNP复合生产实现高效率和低毒性仍然是一个挑战.

研究的目的:

为了在LNP中改进基因组编辑,设计增强的Cas9变体.
评估这些工程 Cas9 RNP-LNP 复合物的有效性,用于各种细胞类型和器官的体内基因编辑.

主要方法:

从Geobacillus stearothermophilus (GeoCas9) 改造出一个耐热的Cas9,并将其改造为iGeoCas9的变种,并增强编辑功能.
配制iGeoCas9与脂质纳米粒子 (LNPs) 的核糖蛋白复合体.
将iGeoCas9 RNP-LNPs静脉注射给记者小鼠,并评估肝脏和肺组织的编辑效率.

主要成果:

与原生GeoCas9.9相比,工程化iGeoCas9变体的基因组编辑效率提高了100倍以上.
iGeoCas9 RNP-LNP复合体成功编辑了各种细胞类型,并诱导了同质导向修复.
在体内研究表明,在一次LNP注射后,小鼠肝脏和肺部的基因组编辑水平为1637%.
通过使用可生物降解的LNP,在肺组织中编辑SFTPC基因的平均效率达到了19%.

结论:

热稳定的Cas9 RNP-LNP复合体为体内基因组编辑提供了显著的进步.
这项技术扩大了基于CRISPR的基因编辑策略的治疗潜力.
与以前的病毒和非病毒传递方法相比,开发的系统显示出更高的效率.

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